CN114391473A - Pumpkin inbred line creation method with white flower mark and excellent comprehensive characters - Google Patents
Pumpkin inbred line creation method with white flower mark and excellent comprehensive characters Download PDFInfo
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- CN114391473A CN114391473A CN202210135894.XA CN202210135894A CN114391473A CN 114391473 A CN114391473 A CN 114391473A CN 202210135894 A CN202210135894 A CN 202210135894A CN 114391473 A CN114391473 A CN 114391473A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
Abstract
The invention provides a pumpkin inbred line creation method with white flower markers and excellent comprehensive properties, which comprises the following steps: hybridizing N87 pumpkin as female parent and wf1 pumpkin as male parent to obtain F1Generation; to F1Selfing to obtain F2Generation, at F2Selecting single white flower plant as male parent to backcross with N87 pumpkin for 3 generations to obtain BC3F1；BC3F1Selfing again to obtain BC3F2(ii) a Selecting BC3F2The white flower in the seed is subjected to single plant selfing to obtain BC3F3(ii) a To BC3F3And carrying out sister crossing in the line to obtain the white flower marked pumpkin inbred line with excellent comprehensive characters. The invention firstly obtains the pumpkin inbred line material with white flower mark and excellent comprehensive characters, overcomes the serious germplasm degeneration phenomenon existing in the traditional technology, and obtains the comprehensive inbred line materialThe characters of the pumpkin hybrid are equivalent to those of the female parent N87, and a feasible way is provided for developing a novel simplified hybrid seed production mode of pumpkin.
Description
Technical Field
The invention belongs to the technical field of agriculture, relates to the field of plant genetic breeding, and particularly relates to a pumpkin inbred line creation method with white flower markers and excellent comprehensive characters.
Disclosure of Invention
Pumpkin (Cucurbita) is one of important vegetable crops, has wide nutritional value, medicinal value and economic value, has the name of 'worldwide vegetables', is the main cultivated species of the pumpkin in China, and can be cultivated in most regions of the country. The Hunan province is the main production area of the Chinese pumpkin, the annual seeding area is more than 70 mu, the Hunan lake melon vegetable production area occupies a dominant position, and the Hunan province plays an important role in the aspects of planting industry structure adjustment, vegetable basket supply protection and the like of the Hunan province.
In the production of hybrid seeds, the flower color is used as a morphological marker, is widely applied to the hybrid seed production of crops, has great advantages in the aspects of improving the seed purity, removing the hybrid plants, identifying the purity of the hybrid seeds and the like, is widely popularized in the hybrid seed production of crops such as rape, rice, cucumber and the like at present, and obviously improves the production efficiency of the hybrid seeds. In the same way, the creation of the pumpkin inbred line material which has the white flower marker character and can be applied to the production of the pumpkin hybrid seeds has important significance for the light simplified hybrid seed production of the pumpkins.
However, the pumpkin flower color is very stable under natural conditions, so that the application of the pumpkin flower color marker to the production of pumpkin hybrids is not reported. More importantly, on the basis, a feasible method is not found out at present how to ensure that the comprehensive properties of the obtained selfing material are not greatly weakened.
In view of the above, there is a need in the art for a method for creating an inbred line of pumpkin having a white flower marker and excellent comprehensive traits.
Aiming at the defects of the prior art, the invention aims to provide a pumpkin inbred line creation method with white flower markers and excellent comprehensive characters.
In order to achieve the purpose, the technical scheme provided by the invention is as follows:
a pumpkin inbred line creation method with white flower markers and excellent comprehensive characters comprises the following steps:
(1) hybridizing N87 pumpkin as female parent and wf1 pumpkin as male parent to obtain F1Generation;
(2) to F1Selfing to obtain F2Generation, at F2Selecting single white flower plant as male parent to backcross with N87 pumpkin for 3 generations to obtain BC3F1；BC3F1Selfing again to obtain BC3F2；
(3) Selecting BC3F2The white flower in the seed is subjected to single plant selfing to obtain BC3F3；
(4) To BC3F3And carrying out sister crossing in the line to obtain the white flower marked pumpkin inbred line with excellent comprehensive characters.
The pumpkin as the male parent material wf1 is a white flower material with stable heredity obtained by EMS mutagenesis, the material has weak plant growth potential, curled leaves, 25 nodes of the first female flower, the average weight of a single pumpkin of 1.90 kg, 21cm of the longitudinal diameter of a fruit, 13cm of the transverse diameter, 2.8cm of pulp thickness, 6.3 percent of soluble solid content, sweet taste, powdery mildew resistance and epidemic disease resistance, and the white flower character is obtained by mutagenesis.
The pumpkin plant of the female parent material N87 has strong growth potential, the main side tendrils can bear the melons, the node position of the first female flower is 20, the average single melon weight is 3.6kg, the longitudinal diameter of the fruit is 27cm, the transverse diameter is 16cm, the pulp thickness is 3.4cm, the soluble solid content is 7.2%, the taste is sweet, and the powdery mildew and epidemic disease resistance are high in field expression.
When the step (2) is carried out, the method specifically comprises the following steps: f is to be1Planting 6 plants in field, building frame for cultivation, and selfing to obtain F2Generation; f is to be260 generations of plants are planted in a field, 6 white flower single plants with comprehensive characters close to N87 are selected as male parents to be backcrossed with female parents N87 to obtain BC1F1Generation; planting BC1F16 plants are bred to obtain BC1F2Generation; mixing BC1F260 generations of plants are planted in a field, 6 white flower single plants with comprehensive characters close to N87 are selected as male parents to be backcrossed with female parents N87 to obtain BC2F1Generation; planting BC2F16 plants are bred to obtain BC2F2Generation; mixing BC2F260 generations of plants are planted in a field, 6 white flower single plants with comprehensive characters close to N87 are selected as male parents to be backcrossed with female parents N87 to obtain BC3F1Generation; planting BC3F16 plants are bred to obtain BC3F2And (4) generation.
When the step (3) is carried out, the concrete steps are as follows: mixing BC3F2Planting 100 plants of 6 generation lines in a field, and strictly selfing and pollinating all white flower single plants in each line; after the fruit is ripe, harvesting all self-bred single melons, performing comprehensive evaluation on all the single melons one by taking the female parent N87 as a control, selecting 6 single melons with the most similar comprehensive properties to the female parent N87 for seed harvest to obtain BC3F3And (4) generation.
When step (4) is performed, BC is added3F3And (3) planting 100 plants of each line of 6 generations in a field, performing mixed pollination in each line, and selectively harvesting 1 pumpkin inbred line material which has white flower marking property and excellent comprehensive property and can be applied to the production of pumpkin hybrid seeds, namely obtaining the white flower marking inbred line material NB-87-1 which is stable, consistent and excellent comprehensive property.
The invention has the beneficial effects that:
1. the invention obtains the inbred line which has the white flower marking property and can be applied to the production of the pumpkin hybrid for the first time, and the male and female flowers of the inbred line are continuous and stable white.
2. The inbred line obtained by the invention has excellent comprehensive properties, and other agronomic properties except the flower color are basically consistent with the core inbred line material N87. Specifically, the inbred line obtained by the invention can normally bloom and bear fruits, the plant growth vigor is strong, the main lateral tendrils can bear melons, the first female flower node is 20, the average single melon weight is 3.4 kg, the longitudinal diameter of the fruit is 27.5cm, the transverse diameter is 16cm, the pulp thickness is 3.5cm, the soluble solid content is 7.1%, the inbred line is sweet in taste, high powdery mildew and epidemic disease resistance is realized in the field, the inbred line can be used as a marked inbred line material to be applied to the production of pumpkin hybrids, and the market blank is filled.
3. The traditional pedigree seed selection needs multi-generation selfing, has serious germplasm degeneration phenomenon, and is difficult to breed the selfing line material with excellent comprehensive characters. The invention has less steps and simple operation; before the inbred line is applied to hybrid production, sister hybridization is carried out in a first generation line, so that the phenomenon of inbreeding degeneration is effectively avoided.
4. The method utilizes the color of the corolla which is visible by naked eyes, convenient to observe, economical and efficient to perform selective pollination, can completely eliminate the interference of non-target materials at one time, greatly lightens the workload and improves the selection efficiency of the character.
Drawings
FIG. 1 is a schematic flow chart of the pumpkin inbred line creation method of the present invention;
FIG. 2 is a photograph of wf1 pumpkin material according to the present invention, wherein part A is a photograph of male flowers and part B is a photograph of female flowers;
FIG. 3 is a photograph of the pumpkin material N87 of the present invention, wherein part A is a photograph of male flower color, part B is a photograph of female flower color, and part C is a photograph of fruit;
FIG. 4 is a photograph of the inbred line obtained by the present invention.
Detailed Description
The present invention is described in detail below by way of examples, and it should be noted that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention.
Interpretation of terms to which the invention relates:
selfing line: a single plant is subjected to continuous multi-generation selfing to form a line.
Selfing: the combination of male and female gametes from the same individual, i.e. the combination of pistil and stamen of the same plant.
And (3) hybridization: the combination of pistils and stamens between plants formed from seeds of two different pumpkins.
Backcrossing: the progeny (including the daughter or daughter generations) are crossed with either of the two parents in a process called backcrossing.
Carrying out sister hybridization: combination of pistils and stamens between different plants in the same line (plants grown from multiple seeds generated by the same pumpkin belong to the same line).
wf 1: using pumpkin high generation comparison line N87 as material, through EMS (ethyl methane sulfonate) mutagenesis, in M2The pumpkin white flower material capable of being stably inherited and obtained by screening in generations is reported in the preliminary analysis of the construction of a Chinese pumpkin mutant library and phenotypic variation (report of nuclear agriculture, 2021, 35(4): 0761-0768). Currently, the breeding is preserved in the pumpkin subject group of vegetable research institute in Hunan province.
N87: the pumpkin subject group of vegetable research institute in Hunan province is an inbred line formed by directionally selecting 4 years and 8 generations of hybrid progeny of a local variety with double peaks in Hunan province and Guangdong candied date pumpkins, wherein the plant growth potential is strong, the 1 st female flower node is about 20, the fruit is in a short gourd shape, the surface of tender fruit is green and has long-strip white spots, the aged fruit is orange red, the surface of the fruit is deep-edged furrows, the taste is powdery and sweet, the content of soluble solids is 7.2%, the average single fruit mass is 3.6kg, and the field shows high powdery mildew and epidemic disease resistance. The breeding of Xin vegetable big fruit honey book (Changjiang vegetables 2015, 12(002), 6-7) of new high-yield, stable-yield and high-quality pumpkin varieties has related reports. Currently, the breeding is preserved in the pumpkin subject group of vegetable research institute in Hunan province.
Example 1
First, hybridization: taking an excellent core inbred line N87 (yellow corolla, strong plant growth potential, capable of bearing melons on main lateral vines, 20 sections of a first female flower node, 3.6 kilograms of average single melon weight, 27cm fruit longitudinal diameter, 16cm transverse diameter, 3.4cm pulp thickness, 7.2 percent of soluble solid content, sweet taste powder and high powdery mildew resistance and epidemic disease resistance expressed in the field) bred by the Hunan vegetable research institute as a female parent, taking a white mutant plant wf1 (white corolla, weak plant growth potential, curled leaves, 25 sections of the first female flower node, 1.90 kilograms of average single melon weight, 21cm fruit longitudinal diameter, 13cm transverse diameter, 2.8cm pulp thickness, 6.3 percent of soluble solid content, sweet taste powder, powdery mildew resistance expressed in the field) obtained by EMS mutagenesis as a male parent, and hybridizing to obtain F1And (4) generation. Obtained F1The generation flower color is normal yellow, and other comprehensive characters are basically between the male parent and the female parent. F1The generation phenotypes were consistent.
Step two, selfing and backcrossing: f is to be1Planting 6 plants in field, building frame for cultivation, and selfing to obtain F2Generation; f is to be260 generations of plants are planted in a field, 6 white flower single plants with comprehensive characters close to N87 are selected as male parents to be backcrossed with female parents N87 to obtain BC1F1Generation; planting BC1F16 plants are bred to obtain BC1F2Generation; mixing BC1F260 generations of plants are planted in a field, 6 white flower single plants with comprehensive characters close to N87 are selected as male parents to be backcrossed with female parents N87 to obtain BC2F1Generation; planting BC2F16 plants are bred to obtain BC2F2Generation; mixing BC2F260 generations of plants are planted in a field, 6 white flower single plants with comprehensive characters close to N87 are selected as male parents to be backcrossed with female parents N87 to obtain BC3F1Generation; planting BC3F16 plants are bred to obtain BC3F2And (4) generation.
Step three, selfing: mixing BC3F2Planting 100 plants of 6 generation lines in a field, and strictly selfing and pollinating all white flower single plants in each line; after the fruit is ripe, harvesting all self-bred single melons, performing comprehensive evaluation on all the single melons one by taking the female parent N87 as a control, selecting 6 single melons with the most similar comprehensive properties to the female parent N87 for seed harvest to obtain BC3F3And (4) generation.
Fourthly, performing sister crossing in the line: mixing BC3F3And (3) planting 100 plants of each line of 6 generations in a field, performing mixed pollination in each line, and selectively harvesting 1 pumpkin inbred line material which has white flower marking property and excellent comprehensive property and can be applied to the production of pumpkin hybrid seeds, namely obtaining the white flower marking inbred line material NB-87-1 which is stable, consistent and excellent comprehensive property.
Experimental example:
the white flower marked inbred line material NB-87-1 with excellent comprehensive properties obtained by breeding by the method of the embodiment 1 is examined and tested.
The male and female flowers are continuously and stably white when planted in the sand in 2021 year, the flowers can normally bloom and bear fruits, the plant growth vigor is strong, the main side tendrils can bear melons, the node position of the first female flower is 20, the average single melon weight is 3.4 kg, the longitudinal diameter of the fruit is 27.5cm, the transverse diameter is 16cm, the pulp thickness is 3.5cm, the soluble solid content is 7.1%, the taste is sweet and powdery, the powdery mildew and epidemic disease resistance is high when the field is expressed, and other agronomic characters except the flower color are basically consistent with the core self-bred material N87. Scanning electron microscope observation is carried out on the male flower crowns of NB-87-1 and N87 on the day of flowering, the epidermal cells of the selfing line NB-87-1 are flat and fuzzy in boundary, the N87 flower crowns are full and clear in boundary, and other characters are not obviously different. The carotenoid content in the inbred lines NB-87-1 and N87 is 0.0617mg/g and 0.0997mg/g respectively; the flavonoid content is 0.3534mg/g/fw and 0.2221mg/g/fw respectively, compared with N87, the carotenoid content of NB-87-1 is reduced by 38.11%, and the flavonoid content is increased by 59.12%.
The white flower material is obtained, and a material foundation is laid for creating the white flower marked inbred line with excellent comprehensive properties. The invention selects the pollination single plant by using the flower color which is visible by naked eyes and easy to observe as the marker character, can completely eliminate the interference of non-target materials at one time, shortens the breeding process and improves the transformation efficiency.
The white flower marked pumpkin inbred line material NB-87-1 bred by the invention has the comprehensive properties reaching the same level as the core inbred line material N87, and provides a feasible way for developing a novel simplified hybrid seed production mode of pumpkin.
Comparative example 1
In the comparative example, EMS is utilized to mutate the high-generation inbred line N87 of the Chinese pumpkin to obtain a white flower plant wf1, female and male flowers of the plant are continuously and stably white, but the plant is weak in growth vigor, curled in leaves, high in node position (25 nodes) of the first female flower, 1.90 kilograms in average single melon weight, 21cm in longitudinal diameter of fruits, 13cm in transverse diameter, 2.8cm in thinner pulp, 7.3% in soluble solid content, sweet in taste, resistant to powdery mildew and epidemic disease, and cannot be directly applied to breeding practice.
Comparative example 2
In the backcross transformation process of the white flower material, the white flower character is transformed into a male parent type high-generation inbred line N219 by the inventor. Transformation processIn the method, N219 is used as a recurrent parent, a fruit type is used as a main selection index, and after 3 generations of backcross, a male parent type white flower material NB-219-1 similar to the fruit type of the N219 is bred. The BC is obtained after NB-219-1 selfing3F2However, BC3F2The single plant of the male parent type white flower in the generation has the conditions of inconsistent fruit types, poor quality and field resistance and the like, and cannot be applied to breeding practice.
Claims (6)
1. A pumpkin inbred line creation method with white flower markers and excellent comprehensive characters is characterized by comprising the following steps:
(1) hybridizing N87 pumpkin as female parent and wf1 pumpkin as male parent to obtain F1Generation;
(2) to F1Selfing to obtain F2Generation, at F2Selecting single white flower plant as male parent to backcross with N87 pumpkin for 3 generations to obtain BC3F1；BC3F1Selfing again to obtain BC3F2；
(3) Selecting BC3F2The white flower in the seed is subjected to single plant selfing to obtain BC3F3；
(4) To BC3F3And carrying out sister crossing in the line to obtain the white flower marked pumpkin inbred line with excellent comprehensive characters.
2. The method of claim 1, wherein said wf1 squash has white corolla, weak plant vigor, curled leaves, and 25 nodes in the first female flower node.
3. The method of claim 1, wherein said N87 squash has yellow crowns, strong plant vigor, can bear melon on both major side vines, and has 20 nodes in the node of the first female flower.
4. The method according to claim 1, wherein step (2) is performed by: f is to be1Planting 6 plants in field, building frame for cultivation, and selfing to obtain F2Generation; f is to be2Planting 60 generation plants in field, selecting 6 white flower single plant zala with most similar comprehensive character to N87 as male parent to backcross with female parent N87 to obtain BC1F1Generation; planting BC1F16 plants are bred to obtain BC1F2Generation; mixing BC1F2Planting 60 generation plants in field, selecting 6 white flower single plant zala with most similar comprehensive character to N87 as male parent to backcross with female parent N87 to obtain BC2F1Generation; planting BC2F16 plants are bred to obtain BC2F2Generation; mixing BC2F2Planting 60 generation plants in field, selecting 6 white flower single plant zala with most similar comprehensive character to N87 as male parent to backcross with female parent N87 to obtain BC3F1Generation; planting BC3F16 plants are bred to obtain BC3F2And (4) generation.
5. The method according to claim 1, wherein step (3) is performed by: mixing BC3F2Planting 100 plants of 6 generation lines in a field, and strictly selfing and pollinating all white flower single plants in each line; after the fruit is ripe, harvesting all self-bred single melons, performing comprehensive evaluation on all the single melons one by taking the female parent N87 as a control, selecting 6 single melons with the most similar comprehensive properties to the female parent N87 for seed harvest to obtain BC3F3And (4) generation.
6. The method of claim 1, wherein step (4) is performed by combining BC3F3And (3) planting 100 plants of each line of 6 generations in a field, performing mixed pollination in each line, and selectively harvesting 1 pumpkin inbred line material which has white flower marking property and excellent comprehensive property and can be applied to the production of pumpkin hybrid seeds to obtain the white flower marked pumpkin inbred line with excellent comprehensive property.
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2022
- 2022-02-15 CN CN202210135894.XA patent/CN114391473B/en active Active
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| US20060123517A1 (en) * | 2004-12-06 | 2006-06-08 | Superak Theodore H | Inbred pumpkin line ZYPMB24 |
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